RNA interference-mediated inhibition of survivin and VEGF in pancreatic cancer cells in vitro

Abstract

The aim of the present study was to investigate the effects of simultaneous short hairpin RNA (shRNA)-targeted survivin and vascular endothelial growth factor (VEGF) inhibition on the proliferation, apoptosis and angiogenesis of human pancreatic cancer cells (Panc-1). Targeted small interfering RNA (siRNA) expression vectors of survivin and VEGF were constructed and transfected into Panc-1 cells. The downregulation of survivin and VEGF expression was evaluated by real-time PCR and western blot analysis. The effects of targeted shRNA on the proliferation and apoptosis of Panc-1 cells were analyzed by MTT assay and flow cytometry (FCM). The culture medium from Panc-1 cells transfected with siRNA was collected and human umbilical vein endothelial cells (HUVECs) were seeded in this media. The proliferation and apoptosis of the HUVECs were also investigated by MTT assay and FCM. A transfected cell line (Panc-1/survivin- shRNA and Panc-1/VEGF-shRNA) was established in which the expression of survivin and VEGF was downregulated. The cell viabilities of Panc-1 cells and HUVECs in the combined inhibition groups were markedly decreased compared with the controls. The cell apoptosis rates of Panc-1 cells and HUVECs in the combined inhibition groups were observed to be significantly increased compared with the controls. The simultaneous RNA interference-mediated downregulation of survivin and VEGF expression inhibited proliferation and induced the apoptosis of Panc-1 cells and HUVECs, indicating that combined therapy with survivin and VEGF inhibition may serve as a potential strategy for the treatment of pancreatic cancer.